Discovery of inhibitors of EBV lytic cycle inducing protein ZTA for therapeutic development

发现 EBV 裂解周期诱导蛋白 ZTA 抑制剂用于治疗开发

• 批准号: 10384443
• 负责人: Takahiro Yano
• 金额: $ 26.91万
• 依托单位: VIRONIKA, LLC
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-16 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10384443
• 关键词: Active Sites; Address; Advanced Development; Affect; Animal Model; Autoimmune Diseases; BZLF1 gene; Binding; Binding Proteins; Biochemical; Biological Assay; Biophysics; Biotechnology; Burkitt Lymphoma; Carcinogens; Cells; Characteristics; Chronic; Collaborations; Communicable Diseases; Computer Models; Computing Methodologies; Crystallization; DNA; DNA Binding; DNA Binding Domain; DNA-Binding Proteins; Detection; Development; Disease; Drug Kinetics; EBV-associated disease; Gene Expression; Genes; Goals; Herpesviridae; Hodgkin Disease; Human; Human Herpesvirus 4; Immune; Immune System Diseases; Immunocompromised Host; Infectious Agent; Infectious Mononucleosis; Intercalating Agents; Lead; Libraries; Life Cycle Stages; Lymphomagenesis; Lytic Phase; Lytic Virus; Malignant Neoplasms; Measures; Medical; Membrane Proteins; Methods; Multiple Sclerosis; Nasopharynx Carcinoma; Natural Products; Non-Hodgkin' s Lymphoma; Nucleotides; Oncogenic; Oxidation-Reduction; Pathology; Periodicity; Persons; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacodynamics; Phase; Population; Production; Proteins; Quantitative Structure-Activity Relationship; Research; Research Project Grants; Risk Factors; Roentgen Rays; Seeds; Small Business Innovation Research Grant; Stomach Carcinoma; Structure; Structure-Activity Relationship; Surface Plasmon Resonance; T-Cell Lymphoma; Testing; Therapeutic Agents; Therapeutic Intervention; Transcription Coactivator; Universities; Validation; Viral; Virus Diseases; base; biophysical techniques; carcinogenesis; cell type; design; detection assay; drug discovery; experience; follow-up; high throughput screening; improved; inhibitor; lead candidate; lead series; lytic gene expression; nanomolar; novel; novel therapeutics; pre-clinical; prevent; programs; screening; skills; small molecule; small molecule inhibitor; small molecule therapeutics; targeted treatment; therapeutic development; tumorigenesis; virus related cancer

Project Summary: The goal of this SBIR is to discover a small molecule inhibitor to the Epstein-Barr Virus (EBV)-encoded ZTA protein to prevent EBV-associated cancer and immune disorders. EBV is a human herpesvirus that switches between latent and lytic gene expression to escape immune detection and promote oncogenic transformation of diverse cell types. EBV is classified as a class I carcinogen by the WHO and estimated to be responsible for ~200,000 new cancer cases per year. EBV-associated cancers are diverse, and include subtypes of gastric carcinoma, nasopharyngeal carcinoma, Burkitt’s lymphoma, Hodgkin’s lymphoma, NK/T cell lymphoma, and various non-Hodgkin’s lymphomas particularly among immunocompromised populations. EBV is the major causative agent of infectious mononucleosis, and the predominant viral risk factor for multiple sclerosis (MS) and other auto immune diseases. The viral-encoded protein, ZTA (also known as BZLF1, ZEBRA, and Z) is a redox-sensitive DNA-binding transcriptional activator that is essential for EBV lytic cycle gene expression and viral production. The periodic cycling of lytic virus is a critical component for EBV-driven malignancy and auto- immune disease, and ZTA and the downstream viral and cellular genes that it activates, have been implicated in carcinogenesis and immune pathology. Therefore, ZTA is an attractive target for therapeutic intervention. The DNA-binding domain of ZTA has been characterized structurally and biochemically, and provides an ideal target for small molecule inhibition of EBV lytic life cycle. We have developed robust biochemical and cell-based assays for detection of ZTA DNA binding and used the biochemical assays for traditional high throughput screening (HTS). We screened the HTS hits using an unrelated DNA binding protein to eliminate DNA intercalators and tested the most selective compounds using surface plasmon resonance (SPR) to determine direct binding to the target protein. We now propose to use improved counter screening assays to characterize the hits from the previous screening efforts, expand our screening to natural products and an additional library of synthetic compounds designed for infectious disease targets. Natural products have advantageous characteristics such as higher rigidity for targeted protein surfaces outside of active sites like that of ZTA. We will use medicinal chemistry to define chemophore structure-activity relationships of our hits and identify a suitable lead compound for further development. Our goal is to produce lead compounds with nanomolar potency in biochemical assays, low micromolar activity in cell-based assays, and greater than 10-fold selectivity against control counter-screens. In Phase 2, we will develop our advanced hits into a pre-clinical lead candidate. The ultimate goal of this SBIR program is to develop a novel small molecule therapeutic agent to block the EBV lytic cycle and its associated diseases.

项目摘要： 该SBIR的目的是发现Epstein-Barr病毒（EBV）编码的ZTA的小分子抑制剂 预防EBV相关癌症和免疫疾病的蛋白质。 EBV是人类的疱疹病毒 在潜在和裂解基因表达之间以逃避免疫检测并促进致癌转化 各种细胞类型。 EBV被WHO归类为I类致癌，估计负责 每年约有200,000例新的癌症病例。 EBV相关的癌症是潜水员，包括胃的亚型 癌，鼻咽癌，伯基特淋巴瘤，霍奇金淋巴瘤，NK/T细胞淋巴瘤和 各种非霍奇金的淋巴瘤，尤其是在免疫功能低下的人群中。 EBV是主要的 感染性单核细胞增多症的病因和多发性硬化症的主要病毒危险因素（MS） 和其他自动免疫疾病。病毒编码的蛋白质ZTA（也称为BZLF1，斑马和Z）是一个 氧化还原敏感的DNA结合转录激活剂对于EBV裂解循环基因表达和 病毒生产。裂解病毒的周期性循环是EBV驱动的恶性肿瘤和自动的关键组成部分 免疫疾病，ZTA以及它激活的下游病毒和细胞基因已被牵涉 在致癌和免疫病理学中。因此，ZTA是治疗干预的有吸引力的目标。 ZTA的DNA结合结构域已在结构和生化上表征，并提供了理想 小分子抑制EBV裂解生命周期的靶标。 我们开发了可靠的生化和基于细胞的测定法，以检测ZTA DNA结合并使用了 传统高通量筛查（HTS）的生化测定。我们使用一个 无关的DNA结合蛋白以消除DNA介导剂并使用最选择性化合物使用 表面等离子体共振（SPR）确定与靶蛋白的直接结合。我们现在建议使用 改进的柜台筛选Assas以表征以前的筛选工作中的命中，扩大我们的 筛选天然产品和为传染病设计的额外的合成化合物库 目标。天然产品具有优势特征，例如靶向蛋白的刚性更高 像ZTA这样的活动站点外面的表面。我们将使用医学化学定义化学成分 命中的结构活性关系，并确定合适的铅化合物以进行进一步发展。我们的 目的是在生化测定中产生具有纳摩尔效力的铅化合物，微摩尔活性低的铅化合物 基于细胞的测定法，对照反屏幕的选择性大于10倍。在第二阶段，我们将 将我们的高级热门歌曲发展为临床前候选人。该SBIR计划的最终目标是 开发一种新型的小分子治疗剂，以阻断EBV裂解周期及其相关疾病。